Proteomic approaches offer distinct advantages in the search for novel ethanol sites of action because they allow large numbers of elements (proteins) to be examined simultaneously in an unbiased fashion. Growing evidence suggests that manipulation of specific protein-protein interactions may provide novel sites to exploit in the search for new drug targets. Identification of such targets will be important for developing improved therapies for complex diseases such as alcoholism. An interaction proteomic approach will be used to identify protein complexes using ethanol sensitive "target" (bait) proteins. The protein targets to be tested in our initial studies were selected because they fulfilled at least two of the following three criteria: (1) they are proteins for which a substantial amount of evidence has accumulated regarding their importance in alcohol actions; (2) they exhibit sensitivity to the effects of alcohol and are being studied by other researchers associated with the P20 application, or (3) they are new proteins identified by microarray screens as possibly playing a role in alcohol actions in vivo. Target proteins include ion channels required for normal synaptic transmission (NMDA receptor (NR1/2B), sodium channel, voltage-gated, type IV, beta (Scn4B)); and, proteins involved membrane trafficking (trafficking protein, kinesin binding 1 (Trakl), potassium channel interacting protein (KChlP3), and synaptotagmins). Interaction proteomics will be used to identify novel protein complexes associated with each of these target proteins in (FVB/NJ x C57BL/6J) hybrid mice. The effect of excessive ethanol consumption (two-bottle choice limited access model) on these protein complexes will be determined. The overall hypothesis is that excessive ethanol consumption alters protein complexes important for normal trafficking and targeting of proteins involved in synaptic transmission.